Mechanical interactions between integrins and extracellular matrix (ECM) are known as critical determinants of cancer and brain disease, however, quantitative analysis of changes in the integrin tension has been limited due to a lack of measurement tools with single molecule precision. To address the critical issue, we have developed a novel DNA-based single molecule force probe, termed tension gauge tether (TGT). With a combination of the TGT assay and micro/nanofabrication techniques, we investigate spatio-temporal changes in integrin tension at the single molecule level during cancer metastasis and amyloidosis. In this seminar, the working principle of the TGT assay will be first introduced and then our new discoveries acquired from the TGT assay will be shown.